Solubilization of pterostilbene and resveratrol for aqueous beverages

ABSTRACT

In one embodiment, the present application discloses compositions and methods of solubilizing pterostilbene or resveratrol, or mixture thereof in aqueous media.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.61/872,971, filed Sep. 3, 2013, which is incorporated herein byreference.

SUMMARY

Pterostilbene (trans-3,5-dimethoxy-4-hydroxystilbene) and resveratrolare natural dietary compounds and the primary antioxidant component ofblueberries, among other fruits. Pterostilbene has increased oralbioavailability in comparison to other stilbene compounds, which mayenhance its dietary benefit and possibly contribute to advantageousclinical effect. Multiple studies have demonstrated the antioxidantactivity of resveratrol and pterostilbene in both in vitro and in vivomodels illustrating both preventative and therapeutic benefits. Theantioxidant activity of resveratrol and pterostilbene has beenimplicated in anticarcinogenesis, modulation of neurological disease,anti-inflammation, attenuation of vascular disease, and amelioration ofdiabetes. Solubility of pterostilbene in aqueous media is very low whichis limiting of its incorporation into beverages for convenientconsumption. We have discovered a methodology to solubilize resveratroland pterostilbene for inclusion in beverages which provides a safe andan efficient delivery of a wide range of resveratrol and pterostilbenedoses.

BACKGROUND

Pterostilbene (trans-3,5-dimethoxy-4-hydroxystilbene) is a naturallyderived compound found in blueberries. The amount of daily pterostilbeneconsumption varies according to dietary fruit intake, and it has beenestimated that pterostilbene content in blueberry varies from 99 ng to520 ng/gram depending on the variety of berry ingested. Substantialevidence suggests that pterostilbene may have numerous preventive andtherapeutic properties in a vast range of human diseases that includeneurological, cardiovascular, metabolic and hematologic disorders (D.McCormack and D. McFadden, 2013, A Review of Pterostilbene Antioxidantand Disease Modification, Oxidative Medicine and Cell Longevity,http://dx.doi.org/10.1155/2013/575482. Further benefits of pterostilbenehave been reported in preclinical trials, in which pterostilbene wasshown to be a potent anticancer agent in several malignancies (D.McCormack and D. McFadden, “Pterostilbene and cancer: current review,”Journal of Surgical Research, vol. 173, no. 2, pp. 53-61, 2012).Pterostilbene is structurally similar to resveratrol, a compound foundin red wine that has comparable antioxidant, anti-inflammatory, andanticarcinogenic properties; however, pterostilbene exhibits increasedabsorption due to the presence of two methoxy groups which cause it toexhibit increased lipophilicity and oral bioavailability. In animalstudies, pterostilbene was shown to have 80% bioavailability compared to20% for resveratrol making it potentially advantageous as a therapeuticagent. The multiple benefits of pterostilbene in the treatment andprevention of human disease have been attributed to its antioxidant,anti-inflammatory, and anticarcinogenic properties leading to improvedfunction of normal cells and inhibition of malignant cell. The evidencereviewed by McCormack and McFadden shows that pterostilbene reducesoxidative stress (OS) and production of reactive oxygen species (ROS),such as hydrogen peroxide (H₂O₂) and superoxide anion (O₂-), which areimplicated in the initiation and pathogenesis of several diseaseprocesses. In addition, various cell lines treated with pterostilbenehave shown increased expression of the antioxidants catalase, totalglutathione (GSH), glutathione peroxidase (GPx), glutathione reductase(GR) and superoxide dismutase (SOD).

Resveratrol (3,5,4′-trihydroxy-trans-stilbene) is a stilbenoid, a typeof natural phenol and a phytoalexin produced naturally by severalplants.

That the many observed benefits of pterostilbene ingestion be realizedfor people, it is necessary that convenient dosage forms be developed.Various beverages are popular with consumers for consumption; sportsdrinks, vitamin waters, fruit juices, energy drinks, etc. However, thenear insolubility of pterostilbene in aqueous solutions precludes theready incorporation of resveratrol or pterostilbene into these popularbeverages.

SUMMARY OF THE INVENTION

In one embodiment, the application discloses a stable and homogeneousaqueous composition comprising: a) a compound selected from the groupconsisting of resveratrol and pterostilbene; and b) an emulsifying agentin an amount sufficient to solubilize resveratrol or pterostilbene toform the stable and homogeneous composition. In another embodiment,there is provided a A stable and homogeneous aqueous formulationcomprising: a) a composition selected from the group consisting of apterostilbene-caffeine co-crystal complex, a resveratrol-caffeineco-crystal complex and a mixture thereof; and b) an emulsifying agent inan amount sufficient to solubilize the pterostilbene-caffeine co-crystalcomplex, a resveratrol-caffeine co-crystal complex and a mixturethereof, to form the stable and homogeneous formulation. In one aspect,the stable and homogeneous aqueous composition further comprisescaffeine. In one aspect, the composition is water soluble. In anotheraspect of the composition, the resveratrol and caffeine is a co-crystalcomplex or the pterostilbene and caffeine is a co-crystal complex. Inanother aspect, the composition further comprises an emulsifying agentselected from the group consisting of TPGS, TPGS-300, TPGS-500,TPGS-600, TPGS-750, TPGS-1000, TPGS-M, TPGS-300-M, TPGS-500-M,TPGS-600-M, TPGS-750-M and TPGS-1000-M, or a mixture thereof. In anotheraspect of the above the composition is an emulsion, a concentratedemulsion or a clear solution in water.

As disclosed herein, the composition is stable, that is, does not resultin decomposition and/or precipitation for a period of at least 6 monthsor 12 months at RT, or at least 12 months at about 12-15° C. Thestability of the solution is also noted where the solution is clear, ornot cloudy or hazy. Clarity may be determined by turbidity units thatmay be measured as Nephelometric Turbidity Units (NTU) as known in theart.

In one embodiment, the present application discloses a method ofsolubilizing resveratrol or pterostilbene, or a mixture of resveratroland pterostilbene, in an aqueous media comprising: a) dissolving theresveratrol or pterostilbene, or a mixture thereof, and an emulsifyingagent in a suitable solvent to form a homogeneous solution; b)evaporating the solvent to form a mixture of resveratrol orpterostilbene, or a mixture thereof, and the emulsifying agent; c)dissolving the resveratrol or pterostilbene mixture, or a mixturethereof, in the aqueous medium by stiffing the mixture with a warmmedium to form a stable emulsion of resveratrol or pterostilbene, or amixture thereof, and emulsifying agent.

In one aspect of the above embodiments, the emulsifying agents are GRAS(Generally Recognized as Safe). In another aspect, the emulsifying agenthas an HLB of greater than 9. In another aspect of each of the above,the emulsifying agent is TPGS (tocopherol polyethylene glycolsuccinate). In another aspect, the ratio of emulsifying agent toresveratrol or pterostilbene, or a mixture thereof, is in the range of3:1 to 10:1. In another aspect of the above, the ratio of emulsifyingagent to resveratrol or pterostilbene, or mixture thereof, is from 4:1to 6:1. In another aspect, the ratio of emulsifying agent to resveratrolor pterostilbene, or a mixture thereof, is from 1:1 to 2:1. In anotheraspect, the resultant emulsion comprises a particle size of less than250 nanometers. In yet another aspect, the resultant emulsion comprisesa particle size of less than 100 nanometers, between 25 and 100nanometers, less than 50 nanometers, or between 15 and 50 nanometers.

In another embodiment, there is provided a method for solubilizingresveratrol or pterostilbene from a co-crystal complex of resveratrol orpterostilbene and caffeine in aqueous media comprising: a) dissolvingthe resveratrol or pterostilbene complex, or a mixture thereof, and anemulsifying agent in a suitable solvent to form a homogeneous solution;b) evaporating the solvent to form a mixture of the resveratrol orpterostilbene complex, or a mixture thereof, and the emulsifying agent;and c) dissolving of the resveratrol or pterostilbene mixture, or amixture thereof, in the aqueous medium by stiffing the mixture with awarm medium to form a stable emulsion of resveratrol or pterostilbene,or a mixture thereof, and emulsifying agent.

In one aspect of the above, the warm medium is provided by heating themixture above RT for a sufficient amount of time to form the stableemulsion. In another aspect, the warm medium is obtained by heating themixture to about 30° C. to about 80° C., about 30° C. to about 70° C.,about 30° C. to about 60° C., about 30° C. to about 50° C., or about 30°C. to about 40° C. In another aspect, the warm medium is obtained byheating the mixture to about 40° C. to about 70° C., or about 40° C. toabout 60° C. In one aspect of the above embodiments, the emulsifyingagents are GRAS (Generally Recognized as Safe). In another aspect, theemulsifying agents have an HLB of greater than 9. In another aspect, theemulsifying agent is TPGS (tocopherol polyethylene glycol succinate),TPGS-300 (D-alpha-tocopheryl polyethylene glycol 300 succinate),TPGS-500, TPGS-600, TPGS-750 and TPGS-1000, or a mixture thereof. Inanother aspect, the emulsifying agent is TPGS-M (tocopherol polyethyleneglycol methyl ether succinate), TPGS-300-M (D-alpha-tocopherylpolyethylene glycol methyl ether 300 succinate), TPGS-500-M, TPGS-600-M,TPGS-750-M and TPGS-1000-M, or a mixture thereof. In another aspect, theemulsifying agent is selected from the group consisting of TPGS,TPGS-300, TPGS-500, TPGS-600, TPGS-750, TPGS-1000, TPGS-M, TPGS-300-M,TPGS-500-M, TPGS-600-M, TPGS-750-M and TPGS-1000-M, or a mixturethereof. In another aspect of the above, the ratio of emulsifying agentto resveratrol or pterostilbene, or a mixture thereof, is in the rangeof 3:1 to 10:1. In another aspect, the ratio of the emulsifying agent toresveratrol or pterostilbene, or a mixture thereof, is 1:1 to 3:1. Inanother aspect, the ratio of the emulsifying agent to resveratrol orpterostilbene, or a mixture thereof, is 4:1 to 5:1. In another aspect,the ratio of the emulsifying agent to resveratrol or pterostilbene, or amixture thereof, is 5:1 to 8:1. In another aspect, the resultantemulsion has a particle size of less than 250 nanometers. In anotheraspect, the resultant emulsion comprises a particle size of less than100 nanometers. In yet another aspect, the resultant emulsion comprisesa particle size of between 25 and 100 nanometers. In another aspect, theresultant emulsion comprises a particle size of below 50 nanometers. Inanother aspect, the resultant emulsion comprises a particle size ofbetween 15 and 50 nanometers. In one aspect of each of the aboveembodiments and aspects, the composition, formulation or emulsioncomprises a mixture of resveratrol and pterostilbene. In one variation,the mixture of resveratrol to pterostilbene is in a ratio of about 95:5,90:10, 85:15, 80:20, 75: 25, 70:30, 65:35, 60: 40, 55:45, 50:50, 45:55,40:60, 35:65, 30:70, 25:75, 20:80, 15:85, 10:90 or 5:95.

In another embodiment of each of the above embodiments and aspect, thereis provided a stable and homogeneous aqueous formulation or compositioncomprising the pterostilbene-caffeine co-crystal complex, aresveratrol-caffeine co-crystal complex and a mixture thereof in wateris at a co-crystal complex:water (wt:wt) ratio of 10:90, 20:80, 30:70,40: 60, 50:50, 60:40, 70:30, 80:20, 90:10, 95:5 or more.

DETAILED DESCRIPTION OF THE INVENTION

We have discovered a methodology to solubilize resveratrol orpterostilbene, or a mixture thereof, such that it may be incorporatedinto aqueous based beverages readily at concentrations sufficient toprovide doses appropriate for provision of the health benefits ofresveratrol or pterostilbene, or a mixture thereof, for consumption.Such beverages include clear beverages, sodas such as Coke® or Pepsi®,fruit juices such as orange juice, apple juice, carbonated ornon-carbonated water or beverages etc . . .

General procedures for preparation of nanoparticulate TPGS basedformulations are found in Arbor Therapeutics, LLC Standard OperatingProcedures; ART 001 Coarse Emulsion Preparation Rev. 1, and ART 003Nicomp 380 ZLS Particle Size Analyses Rev. 1. Exceptions to these SOPsare noted as needed.

Abbreviations: TPGS—D-a-Tocopheryl Polyethyleneglycol-1000-succinate,PSB—Pterostilbene; RVT-Resveratrol; IPA—Isopropanol; REM—resultantemulsion; %-percent of total solids; W/V—weight to volume; Recovery,%-percent of PSB recovered in resultant emulsion after sterile 0.22 μmfiltration; mfg—manufacturing; ND—not determined; BDL—below detectionlimit; ON—overnight.

List of equipment used:

Description Manufacturer Model Number 400 gram balance Denver InstrumentSI-403 100 gram balance Denver Instrument APX-100 Magnetic Barnsteadstirring plate Thermolyne, Cimarec Particle Sizer Particle SizingSystems Nicomp ™ 380 ZLS HPLC Agilent HP 1100 SeriesList of materials used:

Reagent Vendor Part Number Lot Number D-a-Tocopheryl Isochem NA1101040048 Polyethyleneglycol- 1000-succinate Pterostilbene ChromaDex.ASB- 00016996-1206 00016996-100 Resveratrol Sigma-Aldrich R-5010 SigmaNA Distilled Water Kroger Distilled Water NA Grocery Sterile Filters,Fisher  50 mL NA 0.22 μm, PES Scientific 09-741-88 150 mL NA 09-741-01500 mL NA 09-761-107 Cuvettes, Fisher 14 955 125 NA polystyreneScientific 4.5 mL Powerade, Powerade DEC3013CCB3B, Lemon Lime 13:37CT931 Lifewater, SOBE AUG2613, strawberry 0115DL022234 dragonfruitMinute Maid, Minute Maid LTF7W4P, Pink lemonade RTKS4LN Welch's Welch'sNE12K16 17 Tropical Carrot 04:46 C Vitamin water, GLACEAU SEP0913CCC1A,acai-blueberry- 10:21 CT931 pomegranate Perform 02, G series GATORADEOCT3013CT564, 1108TL020235 Pluronic 31R1 Sigma-Aldrich 1001438194MKBC5295V Poly (Propylene 435503 glycol)-block- Poly (Ethyleneglycol)-block-Poly (Propylene glycol) PPG-PEG-PPG Pluronic 17R4Sigma-Aldrich 1001452647 02011BJV Poly (Propylene 435481 glycol)-block-Poly (Ethylene glycol)-block-Poly (Propylene glycol) PPG-PEG-PPGPluronic L-64 Sigma-Aldrich 1001442463 MKBF4936V Poly (Propylene 435449glycol)-block- Poly (Ethylene glycol)-block-Poly (Propylene glycol)PPG-PEG-PPG Pterostilbene and Chromadex 00016988 00016988-0204 CaffeineCo- Crystal Caffeine, 99% Alfa Aesar A10431 B06Y038 Cremophor ® ELPSigma-Aldrich 30906 BCBH0387V

For the preparation of TPGS-750-M and related compounds, see“TPGS-750-M: A Second-Generation Amphiphile for Metal-CatalyzedCross-Couplings in Water at Room Temperature” Lipshutz, B. H. et al. J.Org. Chem. 2011, 76, 4379-4391, and Sigma Aldrich. The followingexperimental description detailed for the analysis of pterostilbene isalso performed similarly with resveratrol, and mixtures thereof. Theanalytical results for resveratrol compositions are similar to thoseobtained for pterostilbene, and a mixture thereof.

Analytical Quantitation of Pterostilbene (PSB) or Resveratrol (RVT).

Quantitation of pterostilbene in concentrated TPGS emulsions employs theanalytical method Pterostilbene.M. or Resveratrol.M. Phenomenex 4.6×50mm Luna 5 82 C18(2) 100A, part number 00B-4252-EO column, mobile phase:isocratic 40/60 acetonitrile/0.01 M H₃PO₄ water for 7 minutes followedby a column wash and re-equilibration, flow rate: 1.5 mL/minute,detection: 254 nm, column temperature: 40° C., and injection volume: 5μL. Sample preparation is a 1:10 dilution with isopropanol.Pterostilbene elutes at 4.5 minutes. A calibration curve/responselinearity was prepared using 0.1, 0.25, 0.5, 1.0 and 2.5 mg/mLsolutions. Response is linear with R² of 0.9999. Since the targetconcentration of pterostilbene in consumer products is expected to beabout 0.08 mg/mL (50-100 mg/drink serving), the sensitivity of themethod was increased by changing the detection wavelength to 310 nm, theabsorbance max of pterostilbene. Quantitation of resveratrol and mixturethereof may be performed as above.

Quantitation of pterostilbene in consumer products employs theanalytical method Pterostilbene 310nm.M. Method: Phenomenex 4.6×50 mmLuna 5 μ C18(2) 100A, part number 00B-4252-EO column, mobile phase:isocratic 40/60 acetonitrile/0.01 M H₃PO₄ water for 7 minutes followedby a column wash and re-equilibration, flow rate: 1.5 mL/minute,detection: 310 nm, column temperature: 40° C., and injection volume: 5μL. Sample preparation is a 1:1 dilution with isopropanol. Pterostilbeneelutes at 4.5 minutes. A calibration curve/response linearity wasprepared using 0.01, 0.025, 0.05, and 0.1 mg/mL solutions. The responseis linear with an R² of 0.9997. Quantitation of resveratrol, or mixturethereof may also employ as above.

Quantitation of pterostilbene and caffeine in one method requires adetection wavelength at which both compounds have UV adsorbance and amobile phase composition change to retain caffeine slightly in theanalysis. Quantitation of pterostilbene and caffeine in concentratedTPGS emulsions employs the analytical method Pterostilbene 280.M.Method: Phenomenex 4.6×50 mm Luna 5 μ C18(2) 100A, part number00B-4252-EO column, mobile phase: 100% 0.01 M H₃PO₄ water for 0.5minutes, 100 to 60% 0.01 M H₃PO₄ water/40% acetonitrile from 0.5 to 2minutes then hold for 5 minutes 40/60 acetonitrile/0.01 M H₃PO₄ waterfollowed by a column wash and re-equilibration, flow rate: 1.5mL/minute, detection: 280 nm, column temperature: 40° C., and injectionvolume: 1 μL. Sample preparation is a 1:10 dilution with isopropanol.Caffeine elutes at 2.4 minutes and pterostilbene elutes at 7.1 minutes.Quantitation of resveratrol and caffeine, and mixture thereof, may usethe above method.

Example 1. Preparation of 5 mg/mL PSB/TPGS, or RVT/TPGS emulsion.

Emulsion preparation: 5.008 g of TPGS was added to 100 mL of distilledwater and mixed on magnetic stiffing plate for 1 hr. 1 g of PSB wasdissolved in 1.2 mL of 95% ethanol. 0.6 mL of ethanol solution of PSBwas added slowly to 100 mL of clear TPGS solution during mixing. Themixture stirred for 2 hr at RT. Upon clearing, the emulsion was filteredthrough 0.22 μm filter, and particle size and PSB content of emulsionnoted in Table 1.

TABLE 1 Particle size, PSB content, and particle stability of resultantemulsion. Formulation Particle Stability Manu- size by PSB Daysfacturing intensity, Content, Recovery, past Particle Date Lot# nm mg/mL% mfg size nm 11 Jun. 2013 005.20.3 23.5 4.08 81.6 14 21.7

HPLC Analysis. “Pterostilbene.M.” method was used for determination ofPSB content. PSB content in resultant emulsion determined by HPLC was4.08 mg/mL. Data indicate that 81.6% of the PSB used for preparation ofthis formulation was incorporated into TPGS particles (Table 1).Analysis of RVT provides similar results with PSB above.

Particle size analysis and stability. The resultant emulsion was stable.Table 1 shows that particle size did not increase over 14 days ofmonitoring. Particle size for RVT are similar to PSB.

Example 2. Preparation of 10 mg/mL PSB/TPGS emulsion. 5.004 g of TPGSand 1.003 g of PSB were weighed and placed in 400 mL beaker. 9 mL of 95%ethanol were added to the solids, and the beaker containing ethanolmixture of TPGS/PSB was placed in the water bath, and incubated at 60°C. for 20 min until clear. The beaker with ethanol TPGS/PSB solution wasplaced into vacuum oven 0/N.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mLof distilled water preheated to 60° C. was added to the highly viscousclear film of TPGS/PBS mixture, the beaker was placed in 60° C. waterbath and total mixture was incubated for 30 min to dissolve. The waterTPGS/PBS mixture was mixed on magnetic stirring plate for 2 hrs at RT.Upon clearing of the mixture, the emulsion was filtered through 0.22 μmfilter, and particle size and PSB content of the resultant emulsion weredetermined (Table 2).

TABLE 2 Particle size, PSB content, and particle stability of resultantemulsion. Formulation Particle Stability Manu- size by PSB Daysfacturing intensity, Content, Recovery, past Particle Date Lot# nm mg/mL% mfg size nm 12 Jun. 2013 005.21.4 26.1 10.57 105.6 40 25.9

HPLC Analysis.“Pterostilbene. 310nm.M.” method was used fordetermination of PSB content. PSB content in resultant emulsiondetermined by HPLC was 10.57 mg/mL. Data indicate that 105.6% of the PSBused for preparation of this formulation was incorporated into TPGSparticles (Table 2). An incorporation value above 100% could resultbecause of water evaporation during 30 min of incubation at 60° C.

Particle size analysis and stability. The resultant emulsion was stable.It can be seen in Table 2 that particle size did not significantlychange over 40 days of monitoring.

Example 3. Dilution of PSB/TPGS emulsion into distilled water.

Procedure 1: PSB/TPGS / Water solution preparation: 170 μl of TPGS/PSBemulsion (lot#005.21.4) was added to 21 mL of Distilled Water. The watersolution of PSB/TPGS emulsion was briefly mixed and stored for 3 days atRT. 3 Days later 0.6 mL aliquot of PSB/TPGS/Water solution was diluted1:1 with IPA, and HPLC analysis of PSB in Table 3a.

TABLE 3a PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.086 ND N/A 0 Jun. 17, 2013 0.086 0.083 97.003

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 3a shows that after 3 days of storage at RT thePSB content in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data shows thatformulated PSB is stable in water.

Procedure 2: PSB/TPGS/Water solution preparation: 170 μl of TPGS/PSBemulsion (lot#005.21.4) was added to 21 mL of Distilled Water. Thesolution of PSB/TPGS emulsion was mixed and stored for 3 and/or 11 daysat RT. 3 or 11 Days later 0.6 mL aliquot of PSB/TPGS / Water solutionwas diluted 1:1 with IPA, filtered through 0.22 μm filter, and HPLCanalysis of PSB content was performed (Table 3b).

TABLE 3b PSB content and Stability of resultant solution. PSB contentCalculated Preparation HPLC PSB, HPLC, Recovery, Days past Date Datemg/mL mg/mL % preparation Jun. 14, 2013 0.086 ND N/A 0 Jun. 17, 20130.086 0.082 95.83 3 Jun. 25, 2013 0.086 0.084 98.17 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 3b shows that after 11 days of storage at RT thePSB content in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the water. Data shows high stabilityof formulated PSB in water.

Procedure 3: PSB/TPGS / Water solution preparation: 170 μl of TPGS/PSBemulsion (lot#005.21.4) was added to 21 mL of Distilled Water. The watersolution of PSB/TPGS emulsion was briefly mixed and stored for 3 days atRT. 3 Days later 1.0 mL aliquot of PSB/TPGS / Water solution wasfiltered through 0.22 μm filter, diluted 1:1 with IPA, and HPLC analysisof PSB content was performed (Table 3c).

TABLE 3c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.086 ND N/A 0 Jun. 17, 2013 0.086 0.073 85.313

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 3c shows that PSB content of the water/PSB/TPGSemulsion when first filtered and then diluted with IPA was significantlylower compared to unfiltered (Procedure 1) and/or dilution with IPAfirst and then filtered (Procedure 2) solutions. Considering that nodegradation of PSB or precipitate was observed in the water, the datasuggest loss of some material on the filter when not diluted with IPA.Procedure 1 and 2 yielded similar PSB content.

Particle size analysis and stability. PSB/TPGS / Water solutionpreparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) was added to 21mL of Distilled Water. The water solution of PSB/TPGS emulsion was mixedand stored for 0, 2, and 6 days at RT. 0, 2, and/or 6 days later 0.3 mLof PSB/TPGS /Water solution was added to the cuvette containing 2.2 mLof distilled water and particle size was measured. To obtain baselinefor distilled water 0.3 mL of distilled water were added to cuvettecontaining 2.2 mL of distilled water. Particle size analysis ispresented in the Table 3d.

TABLE 3d Particle size and stability. Preparation Particle size Dayspast Date Material by intensity, nm preparation Jun. 19, 2013 Water N/A0 Jun. 19, 2013 Water plus Lot# 005.21.4 36.6 0 Jun. 21, 2013 Water plusLot# 005.21.4 37.7 2 Jun. 25, 2013 Water plus Lot# 005.21.4 38.0 6

Particle size did not change over 6 days of storage at RT. Dilution inwater does not impair emulsion stability. Data show chemical andparticle stability of formulated PSB.

Example 4. Preparation of PSB/TPGS/Reb A emulsion. 100 mg of Reb A wasadded directly to 5 mL of PSB/TPGS emulsion lot# 005.21.4 and mixed onmagnetic stiffing plate at RT until clear (˜30 min). Upon clearing ofthe mixture, the emulsion was filtered through 0.22 μm filter, andparticle size and PSB content of the emulsion determined (Table 4).

TABLE 4 Particle size, PSB content, and particle stability of resultantemulsion. Formulation Particle Stability Manu- size by PSB Daysfacturing intensity, Content, Recovery, past Particle Date Lot# nm mg/mL% mfg size nm 14 Jun. 2013 005.21.5 16.2 9.60 90.8 4 16.1

HPLC Analysis. “Pterostilbene.M.” method was used for determination ofPSB content. PSB content in resultant emulsion determined by HPLC was9.6 mg/mL. The data indicate that 90.8% of the PSB determined inemulsion lot # 005.21.4 was recovered in Reb A containing emulsion(Table 4). Direct filtration of the aqueous PSB/TPGS emulsion withoutfirst dilution in IPA causes some loss to the filter by adsorption ontothe filter membrane.

Particle size analysis and stability. The particle size of the Reb Acontaining emulsion was significantly smaller than the emulsion with PSBand TPGS only. The resultant emulsion was stable. As seen in Table 4particle size did not change over 4 days of monitoring.

Example 5. Dilution of PSB/TPGS/Reb A emulsion in distilled water.

Procedure 1: PSB/TPGS/Reb A water solution preparation: 170 μl ofPSB/TPGS/Reb A emulsion (lot#005.21.5) was added to 20 mL of distilledwater. The water solution of PSB/TPGS/Reb A emulsion (lot# 005.21.5) wasmixed and stored for 3 days at RT. 3 Days later 0.6 mL aliquot ofPSB/TPGS/Reb A/Water solution was diluted 1:1 with IPA, and HPLCanalysis of PSB content was performed (Table 5a).

TABLE 5a PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.082 ND N/A 0 Jun. 17, 2013 0.082 0.081 99.263

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 5a shows that after 3 days of storage at RT PSBcontent of resultant solution determined by HPLC was similar tocalculated amount of PSB added to the water. Data shows that TPGS/Reb Aformulated PSB is stable in water.

Procedure 2: PSB/TPGS/Reb A water solution preparation: 170 μl ofPSB/TPGS/Reb A emulsion (lot#005.21.5) was added to 20 mL of distilledwater. The solution of PSB/TPGS/Reb A emulsion was mixed and stored for3 or 11 days at RT. 3 or 11 Days later 0.6 mL aliquot of PSB/TPGS/Reb Awater solution was diluted 1:1 with IPA, filtered through 0.22 μmfilter, and HPLC analysis of PSB content (Table 5b).

TABLE 5b PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.082 ND N/A 0 Jun. 17, 2013 0.082 0.081 99.263 Jun. 25, 2013 0.082 0.083 101.72 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 5b shows that after 11 days of storage at RT PSBcontent in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the aqueous medium. Data suggest thatTPGS/Reb A formulated PSB is stable in water.

Procedure 3: PSB/TPGS/Reb A water solution preparation: 170 μl ofPSB/TPGS/Reb A emulsion (lot#005.21.5) was added to 20 mL of distilledwater. Water solution of PSB/TPGS/Reb A emulsion was briefly mixed andstored for 3 days at RT. 3 Days later 1.0 mL aliquot of PSB/TPGS/RebA/Water solution was filtered through 0.22 μm filter, diluted 1:1 withIPA, and HPLC analysis of PSB content was performed (Table 5c).

TABLE 5c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.082 ND N/A 0 Jun. 17, 2013 0.082 0.068 83.333

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 5c shows that PSB content of the emulsion mixturedirectly filtered and then diluted with IPA was significantly lowercompared to unfiltered (Procedure 1) and/or diluted with IPA first andthen filtered (Procedure 2) solutions. No degradation of PSB orprecipitate was observed in the water, the data rather suggest the lossof some material on the filter when the emulsion preparation is notdiluted with IPA. Procedure 1 and 2 yielded similar PSB content.

Particle size analysis and stability. PSB/TPGS/Reb A/ Water solutionpreparation: 170 μl of PSB/TPGS/Reb A emulsion (lot#005.21.5) was addedto 20 mL of distilled water. Water solution of PSB/TPGS/Reb A emulsionwas briefly mixed and stored for 6 days at RT. Immediately afterpreparation and/or 6 days later 0.3 mL of PSB/TPGS/Reb A/Water solutionwas added to the cuvette containing 2.2 mL of distilled water andparticle size was measured. To obtain baseline for distilled water 0.3mL of distilled water were added to cuvette containing 2.2 mL ofdistilled water. See Table 5d.

TABLE 5d Particle size and stability. Preparation Particle size Dayspast Date Material by intensity, nm preparation Jun. 19, 2013 Water N/A0 Jun. 19, 2013 Water plus Lot# 005.21.5 20.9 0 Jun. 25, 2013 Water plusLot# 005.21.5 21.7 6

The particle size did not change over 6 days of storage at RT. Datasuggest that dilution in water does not impair PSB/TPGS/Reb A emulsion(lot# 005.21.5) stability under these conditions (Table 5d). Data showschemical and particle stability of PSB in water.

Example 6. Preparation of PSB/TPGS/PowerAde solution.

TABLE 6 a Formulation composition. mg per 22 ml Date TPGS PSB PoweradeJun. 14, 2013 8.5 1.7 22 ml

Procedure 1: PSB/TPGS /PowerAde solution preparation: 170 μl of TPGS/PSBemulsion (lot#005.21.4) was added to 22 mL of PowerAde beverage.PowerAde solution of PSB/TPGS emulsion was mixed and stored for 3 daysat RT. 3 Days later 0.6 mL aliquot of PSB/TPGS/PowerAde solution wasdiluted 1:1 with IPA, and HPLC analysis of PSB content was performed(Table 6b).

TABLE 6 b PSB content of resultant solution. Stability. PSB contentCalculated Preparation HPLC PSB, HPLC, Recovery, Days past Date Datemg/ml mg/ml % preparation Jun. 14, 2013 0.082 ND N/A 0 Jun. 17, 20130.082 0.079 96.72 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 6b that after 3 days of storage at RT PSB contentin resultant solution determined by HPLC was similar to calculatedamount of PSB added to the beverage. Data suggest that TPGS formulatedPSB is stable in PowerAde beverage.

Procedure 2: PSB/TPGS/ PowerAde solution preparation: 170 μl of PSB/TPGSemulsion (lot#005.21.4) was added to 22 mL of PowerAde beverage.PSB/TPGS/ PowerAde solution was briefly mixed and stored for 3 or 11days at RT. 3 or 11 Days later 0.6 mL aliquot of PSB/TPGS/PowerAdesolution was diluted 1:1 with IPA, filtered through 0.22 μm filter, andHPLC analysis of PSB content was performed (Table 6c).

TABLE 6c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.082 ND N/A 0 Jun. 17, 2013 0.082 0.079 96.723 Jun. 25, 2013 0.082 0.081 99.17 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 6c shows that after 11 days of storage at RT PSBcontent in the resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data shows that TPGSformulated PSB is stable in PowerAde beverage.

Procedure 3: PSB/TPGS/ PowerAde solution preparation: 170 μl of PSB/TPGSemulsion (lot#005.21.4) was added to 22 mL of PowerAde beverage.PSB/TPGS/ PowerAde solution was briefly mixed and stored for 3 days atRT. 3 Days later 0.6 mL aliquot of PSB/TPGS/PowerAde solution wasfiltered through 0.22 μm filter, diluted 1:1 with IPA, and HPLC analysisof PSB content was performed (Table 6d).

TABLE 6d PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.082 ND N/A 0 Jun. 17, 2013 0.082 0.062 75.913

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 6d (recovery %) shows that PSB content of directlyfiltered PSB/TPGS/PowerAde solution and then dilution with IPA wassignificantly lower compared to unfiltered (Procedure 1) and/or dilutedwith IPA and then filtered (Procedure 2) solutions. PSB recovery infiltered PowerAde solution of formulated PSB was significantly lowercompared to that of filtered PSB/TPGS water solution (examples 3 and 5).Data shows that part of the formulated PSB was bound to the PowerAdecoarse matrix and was retained on the filter during filtration. Dilutionof PSB/TPGS/PowerAde solution with IPA prior to filtration releases PSBfrom the PowerAde matrix and prevents its filter retention.

Particle size analysis and stability. PSB/TPGS/ PowerAde solutionpreparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) was added to 22mL of PowerAde beverage. PSB/TPGS/PowerAde solution was mixed and storedfor 2 and 6 days at RT.

To avoid contribution of coarse beverage matrix to particle sizemeasurement, the beverage was centrifuged at 13,000 rpm for 15 min in aneppendorf centrifuge. To measure particle size of the beverage alone,0.3 mL of supernatant were added to cuvette containing 2.2 mL ofdistilled water. To determine effect of the beverage on particle size ofTPGS/PSP emulsion, 0.6 mL of emulsion (lot # 005.21.4) was mixed with0.6 mL of supernatant, and 0.6 mL of 1:1 mixture was added to thecuvette containing 1.9 mL of distilled water. See Table 6e.

TABLE 6 e Particle size and stability. Particle size by Preparationintensity, Days past Date Material nm preparation Jun. 19, 2013Distilled Water plus Lot# 005.21.4 36.6 0 Jun. 19, 2013 Powerade 609.9 0Jun. 19, 2013 Powerade plus Lot# 005.21.4 40.7 0 Jun. 21, 2013 Poweradeplus Lot# 005.21.4 39.6 2 Jun. 25, 2013 Powerade plus Lot# 005.21.4 41.66

There was 4 nm initial increase of the particle size of the TPGS/PSBemulsion upon dilution in Powerade beverage, the particle size did notchange over next six days. Absence of further particle size changesuggests stability of formulated PSB in Powerade beverage. Data obtainedindicate chemical and particle stability of formulated PSB in Poweradebeverage and suggest 100% of PSB consumption with the beverage.

Example 7. Preparation of PSB/TPGS/LifeWater solution.

TABLE 7a Formulation composition. mg per 21 mL Date TPGS PSB LifeWaterJun. 14, 2013 8.5 1.7 21 mL

Procedure 1: PSB/TPGS/LifeWater solution preparation: 170 μl of TPGS/PSBemulsion (lot#005.21.4) was added to 21 mL of LifeWater beverage.LifeWater solution of PSB/TPGS emulsion was mixed and stored for 3 daysat RT. 3 Days later 0.6 mL aliquot of PSB/TPGS/LifeWater solution wasdiluted 1:1 with IPA, and HPLC analysis of PSB in Table 7b.

TABLE 7b PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation Jun. 14, 2013 0.086 ND N/A 0 Jun. 17, 2013 0.086 0.082 95.833

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 7b shows that after 3 days of storage at RT PSBcontent in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. The data shows that TPGSformulated PSB is stable in LifeWater beverage.

Procedure 2: PSB/TPGS/ LifeWater solution preparation: 170 μl ofPSB/TPGS emulsion (lot#005.21.4) was added to 21 mL of LifeWaterbeverage. PSB/TPGS/LifeWater solution was briefly mixed and stored for 3or 11 days at RT. 3 or 11 Days later 0.6 mL aliquot ofPSB/TPGS/LifeWater solution was diluted 1:1 with IPA, filtered through0.22 μm filter, and HPLC analysis of PSB content was performed (Table7c).

TABLE 7c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.077 89.99 325 Jun. 2013 0.086 0.091 106.35 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 7c shows that after 11 days of storage at RT PSBcontent in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data suggest that TPGSformulated PSB is stable in LifeWater beverage.

Procedure 3: PSB/TPGS/LifeWater solution preparation: 170 μl of PSB/TPGSemulsion (lot#005.21.4) was added to 21 mL of LifeWater beverage.PSB/TPGS/LifeWater solution was briefly mixed and stored for 3 days atRT. 3 Days later 0.6 mL aliquot of PSB/TPGS/LifeWater solution wasfiltered through 0.22 μm filter, diluted 1:1 with IPA, and HPLC analysisof PSB content was performed (Table 7d).

TABLE 7d PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.069 80.64 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 7d (recovery %) shows that PSB content of directlyfiltered PSB/TPGS/LifeWater solution and dilution with IPA was lowercompared to unfiltered (Procedure 1) and/or diluted with IPA and thenfiltered (Procedure 2) solutions. PSB recovery in filtered LifeWatersolution of formulated PSB was similar to that of filtered PSB/TPGSwater solution (examples 3 and 5). Data shows no significant binding offormulated PSB to LifeWater matrix.

Particle size analysis and stability. PSB/TPGS/LifeWater solutionpreparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) was added to 21mL of LifeWater beverage. PSB/TPGS/LifeWater solution was briefly mixedand stored for 2 and 6 days at RT. The beverage was centrifuged at13,000 rpm for 15 min in an eppendorf centrifuge. To measure particlesize of the beverage alone, 0.3 mL of supernatant were added to cuvettecontaining 2.2 mL of distilled water. To determine effect of thebeverage on particle size of TPGS/PSP emulsion, 0.6 mL of emulsion (lot# 005.21.4) was mixed with 0.6 mL of supernatant, and 0.6 mL of 1:1mixture was added to the cuvette containing 1.9 mL of distilled water.See Table 7e.

TABLE 7e Particle size and stability. Particle size by Preparationintensity, Days past Date Material nm preparation 19 Jun. 2013 DistilledWater plus Lot# 005.21.4 36.6 0 19 Jun. 2013 LifeWater 376.9 0 19 Jun.2013 LifeWater plus Lot# 005.21.4 39.2 0 21 Jun. 2013 LifeWater plusLot# 005.21.4 37.9 2 25 Jun. 2013 LifeWater plus Lot# 005.21.4 38.9 6

Although, there was 3 nm initial increase of the particle size of theTPGS/PSB emulsion upon dilution in LifeWater beverage, the particle sizedid not change over next six days of monitoring. Data shows thatcontribution of beverage only particles into final particle size of themixture was not significant. Absence of further particle size changesuggests stability of formulated PSB in LifeWater beverage. Dataindicate chemical and particle stability of formulated PSB in LifeWaterbeverage and show 100% of PSB consumption with the beverage.

Example 8. Preparation of PSB/TPGS/Minute Maid solution.

TABLE 8a Formulation composition. mg per 22 mL Minute Date TPGS PSB Maid14 Jun. 2013 8.5 1.7 22 mL

Procedure 1: PSB/TPGS/Minute Maid solution preparation: 170 μl ofTPGS/PSB emulsion (lot#005.21.4) was added to 22 mL of Minute Maidbeverage. Minute Maid solution of PSB/TPGS emulsion was briefly mixedand stored for 3 days at RT. 3 Days later 0.6 mL aliquot ofPSB/TPGS/Minute Maid solution was diluted 1:1 with IPA. See Table 8b.

TABLE 8b PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.082 ND N/A 0 17 Jun. 2013 0.082 0.075 91.82 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 8b shows that after 3 days of storage at RT PSBcontent was determined by HPLC was similar to calculated amount of PSBadded to the beverage. Data suggest that TPGS formulated PSB is stablein Minute Maid beverage.

Procedure 2: PSB/TPGS/ Minute Maid solution preparation: 170 μl ofPSB/TPGS emulsion (lot#005.21.4) was added to 22 mL of Minute Maidbeverage. PSB/TPGS/Minute Maid solution was briefly mixed and stored for3 or 11 days at room temperature (RT). 3 or 11 Days later 0.6 mL aliquotof PSB/TPGS/Minute Maid solution was diluted 1:1 with IPA, filteredthrough 0.22 μm filter. HPLC analysis of PSB content in Table 8c.

TABLE 8c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.082 ND N/A 0 17 Jun. 2013 0.082 0.077 94.27 325 Jun. 2013 0.082 0.089 108.97 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 8c shows that after 11 days of storage at RT PSBcontent in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data suggest that TPGSformulated PSB is stable in Minute Maid beverage.

Procedure 3: PSB/TPGS/ Minute Maid solution preparation: 170 μl ofPSB/TPGS emulsion (lot#005.21.4) was added to 22 mL of Minute Maidbeverage. PSB/TPGS/Minute Maid solution was briefly mixed and stored for3 days at RT. 3 Days later 0.6 mL aliquot of PSB/TPGS/Minute Maidsolution was filtered through 0.22 μm filter, diluted 1:1 with IPA andHPLC analysis of PSB content was performed (Table 8d).

TABLE 8d PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.082 ND N/A 0 17 Jun. 2013 0.082 0.056 68.56 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 8d (recovery %) shows that PSB content of directlyfiltered PSB/TPGS/Minute Maid solution and then dilution with IPA wassignificantly lower compared to unfiltered (Procedure 1) and/or dilutedwith IPA and then filtered (Procedure 2) solutions. PSB recovery infiltered Minute Maid solution of formulated PSB was notably lowercompared to that of filtered water PSB/TPGS solution (examples 3 and 5).Data suggest that a substantial portion of formulated PSB was bound tothe Minute Maid coarse matrix and was retained on the filter duringfiltration. Dilution of PSB/TPGS/ Minute Maid solution with IPA prior tofiltration releases PSB from the Minute Maid matrix and prevents itsfilter retention.

Particle size analysis and stability. PSB/TPGS/Minute Maid solutionpreparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) was added to 22mL of Minute Maid beverage. PSB/TPGS/Minute Maid solution was brieflymixed and stored for 2 and 6 days at RT. The beverage was centrifuged at13,000 rpm for 15 min in an eppendorf centrifuge. To measure particlesize of the beverage alone, 0.3 mL of supernatant were added to cuvettecontaining 2.2 mL of distilled water. To determine effect of thebeverage on particle size of TPGS/PSP emulsion, 0.6 mL of emulsion (lot# 005.21.4) was mixed with 0.6 mL of supernatant, and 0.6 mL of 1:1mixture was added to the cuvette containing 1.9 mL of distilled water.See Table 8e.

TABLE 8e Particle size and stability. Particle size by Preparationintensity, Days past Date Material nm preparation 19 Jun. 2013 DistilledWater plus Lot# 005.21.4 36.6 0 19 Jun. 2013 Minute Maid 392.2 0 19 Jun.2013 Minute Maid plus Lot# 005.21.4 47.9 0 21 Jun. 2013 Minute Maid plusLot# 005.21.4 47.3 2 25 Jun. 2013 Minute Maid plus Lot# 005.21.4 48.4 6

There was a 11 nm increase of the particle size of formulated PSB upondilution in Minute Maid beverage. The particle size did not change overthe next six days of monitoring. These results suggest significantcontribution of beverage only particles into final particle size of themixture. The light scattering data are in agreement with HPLC datademonstrating substantial binding of formulated PSB to Minute Maidcoarse matrix. Absence of further particle size change suggestsstability of formulation. Data indicate chemical and particle stabilityof formulated PSB in Minute Maid beverage and suggest 100% of PSBconsumption with beverage.

Example 9. Preparation of PSB/TPGS/Welch's Tropical Carrot solution.

TABLE 9a Formulation composition. mg per 20 mL Date TPGS PSB Welch'sTropical Carrot 14 Jun. 2013 8.5 1.7 20 mL

Procedure 1: PSB/TPGS/Welch's Tropical Carrot solution preparation: 170μl of TPGS/PSB emulsion (lot#005.21.4) was added to 20 mL of Welch'sTropical Carrot beverage. Welch's Tropical Carrot solution of PSB/TPGSemulsion was briefly mixed and stored for 3 days at RT. 3 Days later 0.6mL aliquot of PSB/TPGS/Welch's Tropical Carrot solution was diluted 1:1with IPA. HPLC analysis of PSB content was not performed due toprecipitation of the matrix (Table 9b).

TABLE 9b PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.090 ND N/A 0 17 Jun. 2013 0.090 N/A N/A 3

Procedure 2: PSB/TPGS/ Welch's Tropical Carrot solution preparation: 170μl of PSB/TPGS emulsion (lot#005.21.4) was added to 20 mL of Welch'sTropical Carrot beverage. PSB/TPGS/Welch's Tropical Carrot solution wasbriefly mixed and stored for 3 or 11 days at RT. 3 or 11 Days later 0.6mL aliquot of PSB/TPGS/Welch's Tropical Carrot solution was diluted 1:1with IPA, filtered through 0.22 μm filter, and HPLC analysis of PSBcontent was performed (Table 9c).

TABLE 9c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.082 ND N/A 0 17 Jun. 2013 0.090 0.085 94.61 325 Jun. 2013 0.090 0.089 99.06 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 9c shows that after 11 days of storage at RT PSBcontent of resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data shows that TPGSformulated PSB is stable in Welch's Tropical Carrot beverage.

Procedure 3: PSB/TPGS/ Welch's Tropical Carrot solution preparation: 170μl of PSB/TPGS emulsion (lot#005.21.4) was added to 20 mL of Welch'sTropical Carrot beverage. PSB/TPGS/Welch's Tropical Carrot solution wasbriefly mixed and stored for 3 days at RT. 3 Days later 0.6 mL aliquotof PSB/TPGS/Welch's Tropical Carrot solution was filtered through 0.22μm filter, diluted 1:1 with IPA, and HPLC analysis of PSB content wasperformed (Table 9d).

TABLE 9d PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.090 ND N/A 0 17 Jun. 2013 0.090 0.034 37.84 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 9d (recovery %) shows that PSB content of directlyfiltered PSB/TPGS/Welch's Tropical Carrot solution and then dilutionwith IPA was dramatically lower compared to unfiltered (Procedure 1)and/or diluted with IPA and then filtered (Procedure 2) solutions. PSBrecovery in filtered Welch's Tropical Carrot solution of formulated PSBwas more than twice lower compared to that of filtered PSB/TPGS watersolution (examples 3 and 5). Data suggest that a major portion offormulated PSB was bound to Welch's Tropical Carrot coarse matrix andwas retained on the filter during filtration. Dilution ofPSB/TPGS/Welch's Tropical Carrot solution with IPA prior to filtrationreleases PSB from Welch's Tropical Carrot matrix and prevents its filterretention. Welch's Tropical Carrot beverage has precipitation of coarsematrix in its original bottle and written manufacturer instruction“Shake well”.

Particle size analysis and stability. PSB/TPGS/Welch's Tropical Carrotsolution preparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) wasadded to 20 mL of Welch's Tropical Carrot beverage. PSB/TPGS/Welch'sTropical Carrot solution was briefly mixed and stored for 2 and 6 daysat RT. The beverage was centrifuged at 13,000 rpm for 15 min. To measureparticle size of the beverage alone, 0.3 mL of supernatant were added tocuvette containing 2.2 mL of distilled water. To determine effect of thebeverage on particle size of TPGS/PSP emulsion, 0.6 mL of emulsion (lot# 005.21.4) was mixed with 0.6 mL of supernatant, and 0.6 mL of 1:1mixture was added to the cuvette containing 1.9 mL of distilled water.Particle size analysis is presented in the Table 9e.

TABLE 9e Particle size and stability. Particle size by Preparationintensity, Days past Date Material nm preparation 19 Jun. 2013 DistilledWater plus Lot# 005.21.4 36.6 0 19 Jun. 2013 Welch's Tropical Carrot328.7 0 19 Jun. 2013 Welch's Tropical Carrot plus 108.7 0 Lot# 005.21.421 Jun. 2013 Welch's Tropical Carrot plus 79.0 2 Lot# 005.21.4 25 Jun.2013 Welch's Tropical Carrot plus 62.4 6 Lot# 005.21.4

There was increase of the particle size of formulated PSB upon dilutionin Welch's Tropical Carrot beverage form 36.6 to 108.7 nm (Table 9 e).Results suggest contribution of beverage only particles into finalparticle size of the mixture. The light scattering data are in agreementwith HPLC data shows binding of formulated PSB to Welch's TropicalCarrot coarse matrix. Particle size decreased over 6 days from 108.7 to62.4 nm (Table 9e). Possible emulsification of the beverage matrixcomponents in the presence of formulated PSB that could improve drinkingquality of Welch's Tropical Carrot beverage. Data shows chemicalstability of formulated PSB in Welch's Tropical Carrot beverage andsuggest that “shaking well” will provide 100% of PSB consumption withthe beverage.

Example 10. Preparation of PSB/TPGS/Vitamin Water solution.

TABLE 10a Formulation composition. mg per 21 mL Date TPGS PSB VitaminWater 14 Jun. 2013 8.5 1.7 21 mL

Procedure 1: PSB/TPGS /Vitamin Water solution preparation: 170 μl ofTPGS/PSB emulsion (lot#005.21.4) was added to 21 mL of Vitamin Waterbeverage. Vitamin Water solution of PSB/TPGS emulsion was briefly mixedand stored for 3 days at RT. 3 Days later 0.6 mL aliquot ofPSB/TPGS/Vitamin Water solution was diluted 1:1 with IPA, and HPLCanalysis of PSB content was performed (Table 10b).

TABLE 10b PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.082 95.83 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 10b shows that after 3 days of storage at RT PSBcontent in the resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data suggest that TPGSformulated PSB is stable in Vitamin Water beverage.

Procedure 2: PSB/TPGS/ Vitamin Water solution preparation: 170 μl ofPSB/TPGS emulsion (lot#005.21.4) was added to 21 mL of Vitamin Waterbeverage. PSB/TPGS/ Vitamin Water solution was briefly mixed and storedfor 3 or 11 days at RT. 3 or 11 Days later, 0.6 mL aliquot ofPSB/TPGS/Vitamin Water solution was diluted 1:1 with IPA, filteredthrough 0.22 μm filter, and HPLC analysis of PSB content was performed(Table 10c).

TABLE 10c PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.082 95.83 325 Jun. 2013 0.086 0.087 101.68 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 10c shows that after 11 days of storage at RT PSBcontent in the resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data suggest that TPGSformulated PSB is stable in Vitamin Water beverage.

Procedure 3: PSB/TPGS/Vitamin Water solution preparation: 170 μl ofPSB/TPGS emulsion (lot#005.21.4) was added to 21 mL of Vitamin Waterbeverage. PSB/TPGS/Vitamin Water solution was briefly mixed and storedfor 3 days at RT. 3 Days later 0.6 mL aliquot of PSB/TPGS/Vitamin Watersolution was filtered through 0.22 μm filter, diluted 1:1 with IPA, andHPLC analysis of PSB content was performed (Table 10d).

TABLE 10 d PSB content of resultant solution. Stability. PSB contentCalculated Preparation HPLC PSB, HPLC, Recovery, Days past Date Datemg/mL mg/mL % preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.0860.070 81.81 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 10d (recovery %) shows that PSB content offiltered PSB/TPGS/Vitamin Water solution and then diluted with IPA wassignificantly lower compared to unfiltered (Procedure 1) and/or dilutedwith IPA and then filtered (Procedure 2) solutions. PSB recovery infiltered Vitamin Water solution of formulated PSB was similar to that offiltered PSB/TPGS water solution (examples 3 and 5). The data provide noevidence of significant binding of formulated PSB to Vitamin Watermatrix.

Particle size analysis and stability. PSB/TPGS/Vitamin Water solutionpreparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) was added to 21mL of Vitamin Water beverage. PSB/TPGS/Vitamin Water solution wasbriefly mixed and stored for 2 and 6 days at RT. To avoid contributionof coarse beverage matrix to particle size measurement, the beverage wascentrifuged at 13,000 rpm for 15 min in an eppendorf centrifuge. Tomeasure particle size of the beverage alone, 0.3 mL of supernatant wereadded to cuvette containing 2.2 mL of distilled water. To determineeffect of the beverage on particle size of TPGS/PSP emulsion, 0.6 mL ofemulsion (lot # 005.21.4) was mixed with 0.6 mL of supernatant, and 0.6mL of 1:1 mixture was added to the cuvette containing 1.9 mL ofdistilled water. Particle size analysis is presented in the Table 10e.

TABLE 10e Particle size and stability. Particle size Days pastPreparation by intensity, prepar- Date Material nm ation 19 Jun. 2013Distilled Water plus Lot# 005.21.4 36.6 0 19 Jun. 2013 Vitamin Water BDL0 19 Jun. 2013 Vitamin Water plus Lot# 005.21.4 41.4 0 21 Jun. 2013Vitamin Water plus Lot# 005.21.4 39.4 2 25 Jun. 2013 Vitamin Water plusLot# 005.21.4 41 6

Although, there was ˜5 nm initial increase of the particle size of theTPGS/PSB emulsion upon dilution in Vitamin Water beverage, the particlesize did not change over 6 days. Data suggest that the contribution ofbeverage only particles into final particle size of the mixture was notsignificant. Absence of further particle size change suggests stabilityof formulated PSB in Vitamin Water beverage. Data obtained indicatechemical and particle stability of formulated PSB in Vitamin Waterbeverage and suggest 100% of PSB consumption with the beverage.

Example 11. Preparation of PSB/TPGS/Gatorade Perform 02 solution.

TABLE 11a Formulation composition. mg per 21 mL Date TPGS PSB GatoradePerform 02 14 Jun. 2013 8.5 1.7 21 mL

Procedure 1: PSB/TPGS/Gatorade Perform 02 solution preparation: 170 μlof TPGS/PSB emulsion (lot#005.21.4) was added to 21 mL of GatoradePerform 02 beverage. Gatorade Perform 02 solution of PSB/TPGS emulsionwas briefly mixed and stored for 3 days at RT. 3 Days later 0.6 mLaliquot of PSB/TPGS/Gatorade Perform 02 solution was diluted 1:1 withIPA, and HPLC analysis of PSB content was performed (Table 11b).

TABLE 11b PSB content of resultant solution. PSB content CalculatedPreparation PSB, HPLC, Recovery, Days past Date HPLC Date mg/mL mg/mL %preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.081 94.66 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 4b shows that after 3 days of storage at RT PSBcontent in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data suggest that TPGSformulated PSB is stable in Gatorade Perform 02 beverage.

Procedure 2: PSB/TPGS/Gatorade Perform 02 solution preparation: 170 μlof PSB/TPGS emulsion (lot#005.21.4) was added to 21 mL of GatoradePerform 02 beverage. PSB/TPGS/Gatorade Perform 02 solution was brieflymixed and stored for 3 or 11 days at RT. 3 or 11 Days later 0.6 mLaliquot of PSB/TPGS/Gatorade Perform 02 solution was diluted 1:1 withIPA, filtered through 0.22 μm filter; HPLC analysis of PSB content wasperformed (Table 11c).

TABLE 11c PSB content of resultant solution. PSB content Re- Days pastPreparation HPLC Calculated HPLC, covery, prepar- Date Date PSB, mg/mLmg/mL % ation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.085 99.343 25 Jun. 2013 0.086 0.085 99.34 11

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 11c shows that after 11 days of storage at RT PSBcontent in resultant solution determined by HPLC was similar tocalculated amount of PSB added to the beverage. Data suggest that TPGSformulated PSB is stable in Gatorade Perform 02 beverage.

Procedure 3: PSB/TPGS/ Gatorade Perform 02 solution preparation: 170 μlof PSB/TPGS emulsion (lot#005.21.4) was added to 21 mL of GatoradePerform 02 beverage. PSB/TPGS/Gatorade Perform 02 solution was brieflymixed and stored for 3 days at RT. 3 Days later 0.6 mL aliquot ofPSB/TPGS/Gatorade Perform 02 solution was filtered through 0.22 μmfilter, diluted 1:1 with IPA, and HPLC analysis of PSB content wasperformed (Table 11d).

TABLE 11d PSB content of resultant solution. PSB content CalculatedPreparation HPLC PSB, HPLC, Recovery, Days past Date Date mg/mL mg/mL %preparation 14 Jun. 2013 0.086 ND N/A 0 17 Jun. 2013 0.086 0.071 82.98 3

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 11d (recovery %) shows that PSB content ofdirectly filtered PSB/TPGS/Gatorade Perform 02 solution and thendilution with IPA was significantly lower compared to unfiltered(Procedure 1) and/or diluted with IPA and then filtered (Procedure 2)solutions. PSB recovery in filtered Gatorade Perform 02 solution offormulated PSB was similar to that of filtered PSB/TPGS water solution(examples 3 and 5). No evidence of significant binding of formulated PSBto Gatorade Perform 02 matrix.

Particle size analysis and stability. PSB/TPGS/ Gatorade Perform 02solution preparation: 170 μl of PSB/TPGS emulsion (lot#005.21.4) wasadded to 21 mL of Gatorade Perform 02 beverage. PSB/TPGS/GatoradePerform 02 solution was briefly mixed and stored for 2 and 6 days at RT.To avoid contribution of coarse beverage matrix to particle sizemeasurement, the beverage was centrifuged at 13,000 rpm for 15 min in aneppendorf centrifuge. To measure particle size of the beverage alone,0.3 mL of supernatant were added to cuvette containing 2.2 mL ofdistilled water. To determine effect of the beverage on particle size ofTPGS/PSP emulsion, 0.6 mL of emulsion (lot # 005.21.4) was mixed with0.6 mL of supernatant, and 0.6 mL of 1:1 mixture was added to thecuvette containing 1.9 mL of distilled water. Particle size analysis ispresented in the Table 11e.

TABLE 11e Particle size and stability. Particle size Days pastPreparation by intensity, prepar- Date Material nm ation 19 Jun. 2013Distilled Water plus 36.6 0 Lot# 005.21.4 19 Jun. 2013 Gatorade Perform02 512.8 0 19 Jun. 2013 Gatorade Perform 02 plus 41 0 Lot# 005.21.4 21Jun. 2013 Gatorade Perform 02 plus 39.4 2 Lot# 005.21.4 25 Jun. 2013Gatorade Perform 02 plus 40.9 6 Lot# 005.21.4

There was 4.4 nm initial increase of the particle size of the TPGS/PSBemulsion upon dilution in Gatorade Perform 02 beverage. The particlesize did not change over next six days of monitoring. Data suggest thatthe contribution of beverage only particles to final particle size ofthe mixture was not significant. Absence of further particle size changesuggests stability of formulated PSB in Gatorade Perform 02 beverage.Data indicates chemical and particle stability of formulated PSB inGatorade Perform 02 beverage and suggest 100% of PSB consumption withthe beverage.

Example 12. Preparation of PSB/TPGS aqueous emulsion. Determine thelowest TPGS/PSB ratio producing stable emulsion containing 10 mg/mL ofPSB in aqueous media.

TABLE 12 a Formulation composition. Weighed, mg (per 100 ml) TPGS/PSBDate Lot# Material TPGS PSB Ratio 01 Jul. 2013 005.30.3 Mixture 31171022 3.05 01 Jul. 2013 005.30.4 Mixture 4040 1044 3.87

Table 12a: Amounts of TPGS and PSB were weighed and placed in 400 mLbeaker. 9 mL of 95% ethanol were added to the solids, and the beakercontaining ethanol mixture of TPGS/PSB was incubated at 60° C. for 20min until clear. and placed into vacuum oven 0/N.

PSB/TPGS emulsions lot# 005.30.7 and lot# 005.30.8 were prepared on 3Jul. 2013. Emulsion preparation: The beaker was removed from vacuum ovenand 100 mL of distilled water preheated to 60° C. was added to thehighly viscous clear film of TPGS/PBS mixture, the beaker was placed ina 60° C. water bath and total mixture was incubated for 30 min tofacilitate dissolution. The water TPGS/PBS mixture was mixed on magneticstiffing plate for 2 hrs at RT. Upon clearing of the mixture, theemulsion was filtered through 0.22 μm filter, and particle size and PSBcontent of the resultant emulsions were determined (Table 12b).

TABLE 12b Particle size, PSB content, and particle stability ofresultant emulsions. Formulation Particle Stability Manu- size by PSBDays Particle facturing intensity, Content, Recovery, past size DateLot# nm mg/mL % mfg nm 3 Jul. 2013 005.30.7 180.4 8.7 85.3 19 216.4 3Jul. 2013 005.30.8 58.0 10.10 96.7 19 60.6

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 12b shows that 4/1 (TPGS/PSB) ratio was the lowestratio that allowed formulation of 10mg/mL of PSB into aqueous media.

Particle size analysis and stability. The resultant emulsion lot #005.30.8 was stable. It can be seen in Table 12b that particle size didnot significantly change over 19 days of monitoring. In contrast,emulsion lot# 005.30.7 was not stable (Table 12b) and formed aprecipitate over the time of monitoring.

Example 13. Preparation of PSB/ PPG-PEG-PPG, Pluronic emulsions. Toinvestigate the possibility of using alternative emulsifying agents,PPG-PEG-PPG, Pluronic block polymers with 2-7, 7-12, and 12-18 HLB(Hydrophilic-lipophilic balance) range to generate PSB containing stableemulsions containing ˜10 mg/mL of PSB. Formulation composition ispresented in the Table 13a.

TABLE 13a Formulation composition. Weighed, mg Block (per 100 mL)polymer/ HLB Block PSB Date Lot# Block polymers Range Material polymerPSB Ratio 11 Jul. 2013 005.32.1.27 PPG-PEG-PPG, 2-7 Mixture 2003 10581.89 Pluronic 31R1 11 Jul. 2013 005.32.2.27 PPG-PEG-PPG, 2-7 Mixture4013 1014 3.96 Pluronic 31R1 11 Jul. 2013 005.32.3.712 PPG-PEG-PPG, 7-12 Mixture 2036 1017 2.0 Pluronic 17R4 11 Jul. 2013 005.32.4.712PPG-PEG-PPG,  7-12 Mixture 3996 1037 3.85 Pluronic 17R4 11 Jul. 2013005.32.5.1218 PPG-PEG-PPG, 12-18 Mixture 1994 1014 1.97 Pluronic L-64 11Jul. 2013 005.32.6.1218 PPG-PEG-PPG, 12-18 Mixture 4007 1013 3.96Pluronic L-64

Table 13a shows amounts of block polymers and PSB were weighed andplaced in 400 mL beaker. 9 mL of 95% ethanol were added to the solids,and the beaker containing ethanol mixture of TPGS/PSB was placed in thewater bath and incubated at 60° C. for 20 min until clear. The beakerwith ethanol TPGS/PSB solution was placed into vacuum oven 0/N.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mLof distilled water preheated to 60° C. was added to the highly viscousclear film of Block polymer/PBS mixture; the beaker was placed in a 60°C. water bath and total mixture was incubated for 30 min to facilitatedissolution. The water Block polymer/PBS mixture was mixed on magneticstiffing plate for 2 hrs at RT. Two hours later the emulsion wasfiltered through 0.22 μm filter, and particle size and PSB content ofthe resultant emulsions were determined (Table 13b).

TABLE 13b Particle size, PSB content, and particle stability ofresultant emulsions. Formulation Particle Stability size by PSB DaysParticle Manufacturing intensity, Content, Recovery, past size Date Lot#nm mg/mL % mfg nm 12 Jul. 2013 005.32.1.27 Not Detectable BDL NA NA NA12 Jul. 2013 005.32.2.27 Not Detectable BDL NA NA NA 12 Jul. 2013005.32.3.712 Not Detectable BDL NA NA NA 12 Jul. 2013 005.32.4.712 NotDetectable BDL NA NA NA 12 Jul. 2013 005.32.5.1218 Not Detectable BDL NANA NA 12 Jul. 2013 005.32.6.1218 6907.9 0.23 2.3 NA NA

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 13b shows that PSB was only detected in Lot#005.32.6.1218 where PPG-PEG-PPG, Pluronic L-64 (HLB Range 12-18) wasused for formulation of PSB. However, the measured PSB content(0.23mg/mL) was far from the target concentration of 10 mg/mL.

Particle size analysis and stability. Table 13b shows that particle sizecould be measured only in lot# 005.32.6.1218 where PPG-PEG-PPG, PluronicL-64 (HLB Range 12-18) was used for formulation of PSB. Emulsion wasunstable and formed a precipitate.

Example 14. Preparation of PSB/ PPG-PEG-PPG, Pluronic emulsions. Toinvestigate the possibility of using different PPG-PEG-PPG, Pluronicblock polymers with 2-7, 7-12, and 12-18 HLB (Hydrophilic-lipophilicbalance) range at higher block polymer/PSB ratio to generate PSBcontaining stable emulsions containing ˜10 mg/mL of PSB.

PSB/ PPG-PEG-PPG, Pluronic block polymers ethanol mixtures were preparedon 16 Jul. 2013. Formulation composition is presented in the Table 14a.

TABLE 14a Formulation composition. Weighed, mg Block (per 100 mL)polymer/ HLB Block PSB Date Lot# Block polymers Range Material polymerPSB Ratio 16 Jul. 2013 005.33.7.27 PPG-PEG-PPG, 2-7 Mixture 10205 100510.15 Pluronic 31R1 16 Jul. 2013 005.33.8.712 PPG-PEG-PPG, 2-7 Mixture10191 1006 10.13 Pluronic 17R4 16 Jul. 2013 005.33.9.1218 PPG-PEG-PPG,Mixture 10731 1003 10.70 Pluronic L-64  7-12

Table 14a shows the amount of block polymers and PSB were weighed andplaced in 400 mL beaker. 9 mL of 95% ethanol were added, and the beakercontaining ethanol mixture of TPGS/PSB was placed in the water bath andincubated at 60° C. for 20 min until clear. The beaker with ethanolTPGS/PSB solution was placed into vacuum oven 0/N.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mLof distilled water preheated to 60° C. was added to highly viscous clearfilm of Block polymer /PBS mixture, beaker was placed to 60° C. waterbath and total mixture was incubated for 30 min to facilitate thedissolution. Water Block polymer /PBS mixture was mixed on magneticstiffing plate for 2 hrs at RT. Two hours later the emulsion wasfiltered through 0.22 μm filter, and particle size and PSB content ofthe resultant emulsions were determined (Table 14b).

TABLE 14b Particle size, PSB content, and particle stability ofresultant emulsions. ART-207 Manufacturing Particle size Content, DateLot# by intensity, nm mg/mL Recovery, % 17 Jul. 2013 005.33.7.27 20576BDL NA 17 Jul. 2013 005.33.8.712 420 0.13 1.3 17 Jul. 2013 005.33.9.1218113 9.47 94.4

HPLC Analysis. “Pterostilbene 310nm.M” method was used for determinationof PSB content. Table 14b shows that PSB was detected in Lot#005.33.8.712 (0.13 mg/mL) where PPG-PEG-PPG, Pluronic 17R4 (HLB Range7-12) and Lot# 005.33.9.1218 (9.47 mg/mL) where PPG-PEG-PPG, PluronicL-64 (HLB Range 12-18) were used for formulation of PSB.

Particle size analysis and stability. Table 14b shows that particle sizedecreased from 20576 to 113 nm with increasing of HLB range. However,emulsion Lot# 005.33.9.1218 was stable only at 40-50° C. and formed aprecipitate at RT. Data obtained demonstrate uniqueness of TPGS(HLB=14.3) as an efficient formulating agent for generating stableemulsions containing 10 mg/mL of PSB in aqueous solutions. Blockpolymers, including PPG-PEG-PPG, Pluronic L-64 with similar HLBproperties to TPGS, were used in an attempt to formulate PSB. None ofthese formulating agents were able to produce stable 10 mg/mL PSBemulsion at similar or even higher than TPGS/PSB ratios.

Experiment 15. Preparation of PSB/Caffeine/TPGS emulsion. To prepareemulsion containing 10 mg/ml of PSB/Caffeine (56.2% w/w PSB and 45.0%w/w Caffeine)

TABLE 15a Formulation composition. Weighed, mg Calculated (per 100 ml)Weight, mg PSB/ based on C of A Date Lot# Material TPGS caffeine PSBCaffeine 30 Jul. 2013 005.35.1 Mixture 5073 1022 574 460

5.073 g of TPGS and 1.022 g of PSB/caffeine were weighed and placed in400 ml beaker. 10 ml of 95% ethanol were added to the solids, and thebeaker containing ethanol mixture of TPGS/PSB/caffeine was placed in thewater bath, and incubated at 60° C. for 20 min until clear. Then beakerwith ethanol TPGS/PSB solution was placed into vacuum oven 0/N.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mlof distilled water was added to highly viscous cloudy film ofTPGS/PSB/Caffeine mixture, the beaker was placed into a 60° C. waterbath and the total mixture was incubated for 30 min to facilitatedissolution. The water TPGS/PSB/Caffeine mixture was mixed on magneticstiffing plate for 2 hrs at RT. Upon clearing of the mixture, particlesize and PSB and caffeine content of the resultant emulsion wasdetermined (Table 15b).

TABLE 15b Particle size, PSB content, and Caffeine content of resultantemulsion. PSB Caffeine Manufacturing Particle size by Content, Content,Date Lot# intensity, nm mg/ml Recovery, % mg/ml Recovery, % 31 Jul. 2013005.35.3 13.1 5.66 99 4.53 98

HPLC Analysis. “Pterostilbene 280.M.” method was used for determinationof PSB and caffeine content. PSB content and Caffeine content inresultant emulsion determined by HPLC (“Pterostilbene 280 NM” method)was PSB was 5.66 mg/mL and caffeine 4.53 mg/mL. Recovery is based on theweighed amount of the pterostilbene/caffeine co-crystal and the weight %provided by the Chromadex C of A. Data indicate that 99% of the PSB usedfor preparation of this formulation was incorporated into TPGS particles(Table 15b).

TABLE 15c Particle size, PSB content, and stability of resultantemulsion. Particle Days size by PSB Caffeine Manufacturing pastintensity, Content, Recovery, Content, Recovery, Date Lot# mfg nm mg/ml% mg/ml % 31 Jul. 2013 005.35.3 6 13.1 5.85 102 4.68 102

Particle size analysis and stability. The ratio of TPGS to PSB in thisemulsion is 8.8:1 and this large ratio produces the small particle sizeof 13.1 nm. The resultant emulsion was stable. Tables 15b and 15c showthat particle size did not change over 6 days of monitoring. After 30days the emulsion is visually unchanged.

Experiment 16. Preparation of PSB/Caffeine/TPGS emulsion. To prepareemulsion containing 10 mg/ml of PSB from the PSB/Caffeine co-crystal(56.2% w/w PSB and 45.0% w/w Caffeine).

TABLE 16a Formulation composition. Weighed, mg Calculated (per 100 ml)Weight, mg PSB/ based on C of A Date Lot# Material TPGS caffeine PSBCaffeine 1 Aug. 2013 005.36.2 Mixture 5018 1779 1000 801

5.018 g of TPGS and 1.779 g of PSB/caffeine were weighed and placed in400 ml beaker. 10 ml of 95% ethanol were added to the solids, and thebeaker containing ethanol mixture of TPGS/PSB/caffeine was placed in thewater bath, and incubated at 60° C. for 20 min until clear. Beaker withethanol TPGS/PSB solution was placed into vacuum oven O/N.

Emulsion preparation: The beaker was removed from oven and 100 ml ofdistilled water was added to highly viscous cloudy film ofTPGS/PSB/Caffeine mixture, beaker was placed to 60° C. water bath andtotal mixture was incubated for 30 min to facilitate the dissolution.Then water TPGS/PSB/Caffeine mixture was mixed on magnetic stiffingplate for 2 hrs at RT. Upon clearing of the mixture, particle size, PSBand caffeine content of the resultant emulsion was determined (Table16b). The emulsion is transparent but hazy. The emulsion was filteredthrough 0.22 μm filter and particle size, PSB and caffeine content weremeasured again. The haziness did not change after filtration.

TABLE 16b Particle size, PSB content, and Caffeine content of resultantemulsion. PSB Caffeine Manufacturing Particle size by Content, Content,Date Lot# intensity, nm mg/ml Recovery, % mg/ml Recovery, % 2 Aug. 2013005.36.3 33.4 9.65 97 7.67 96 2 Aug. 2013 005.36.3 33.5 9.75 98 7.73 97filtered

HPLC Analysis. “Pterostilbene 280.M.” method was used for determinationof PSB and caffeine content. PSB content and Caffeine content inresultant emulsion determined by HPLC (“Pterostilbene 280 NM” method)was PSB was 9.65 mg/mL and caffeine 7.67 mg/mL. Particle size, PSB andcaffeine content are not significantly different in the unfilteredemulsion as compared to the filtered emulsion. Recovery is based on theweighed amount of the PSB/caffeine co-crystal and the weight % providedby the Chromadex C of A. Data show 97% of the PSB used for preparationwas incorporated into TPGS particles (Table 16b).

Particle size analysis. The ratio of TPGS to PSB in this emulsion is 5:1and this ratio resulted in a particle size of 33.4 nm which is largerthan the particle size in Experiment 15 and shows that a lower ratio ofPSB and TPGS results in larger particle sizes. The presence of caffeinedoes not appear to influence the emulsification of PSB. Three days aftermanufacture, the filtered emulsion was found to have crystallized. Theanalytical data of the supernatant is shown in Table 16c below. Both PSBand Caffeine have precipitated from solution. The ratio of TPGS to PSBin the supernatant or mother liquors is 9.1:1. The results for particlesize and PSB and caffeine content of the supernatant suggest the maximumstable concentrations of PSB and caffeine and minimum ratio of PSB toTPGS in an emulsion for a given concentration of TPGS (50 mg/mL). Theratio of TPGS to PSB in the supernatant is 9.1:1.

TABLE 16c Particle size, PSB content, and stability of resultantemulsion. Particle Days size by PSB Caffeine Manufacturing pastintensity, Content, Recovery, Content, Recovery, Date Lot# mfg nm mg/ml% mg/ml % 2 Aug. 2013 005.36.3 3* 13.8 5.49 55 4.58 57 filtered *theemulsion crystallized, analysis is of the supernatant.

Dilutions. On 9Aug. 2013 (one week after preparation) Lot# 005.36.3 waswarmed to 65° C. for 1.5 hours. The crystals had dissolved and thesolution was cloudy. Upon cooling and setting for 2 hours on the bench,the solution cleared and two phases were observed. Vigorous handagitation for 15 seconds resulted in a solution which appeared to behomogenous. The particle size analysis result of the solution with thecrystals re-dissolved was 31.5 nm. Dilutions of the preparation weremade to evaluate the effect of concentration of the PSB and caffeine onemulsion stability. Dilutions: none, 1:1, 1:10, 1:20 and 1:100 wereprepared in 50 mL centrifuge tubes and allowed to set for three days. Onday three the no dilution sample had crystallized and the 1:1 dilutionsample had a few crystals. The other dilutions had no crystals. Resultssuggest that when the PSB concentration is below 0.6 mg/mL and thecaffeine concentration similarly low (below 0.5 mg/mL), a stableemulsion can be readily generated.

Experiment 17. Preparation of PSB/Caffeine/TPGS emulsion. To prepareemulsion containing 11.2 mg/ml of PSB from the PSB/Caffeine co-crystal(56.2% w/w PSB and 45.0% w/w Caffeine).

TABLE 17a Formulation composition. Weighed, mg Calculated (per 100 ml)Weight, mg PSB/ based on C of A Date Lot# Material TPGS caffeine PSBCaffeine 1 Aug. 2013 005.36.1 Mixture 5003 2.008 1128 904

5.003 g of TPGS and 2.008 g of PSB/caffeine were weighed and placed in400 ml beaker. 10 ml of 95% ethanol were added to the solids, and thebeaker containing ethanol mixture of TPGS/PSB/caffeine was placed in thewater bath, and incubated at 60° C. for 20 min until clear. Then beakerwith ethanol TPGS/PSB solution was placed into vacuum oven 0/N.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mlof distilled water was added to highly viscous cloudy film ofTPGS/PSB/Caffeine mixture, beaker was placed to 60° C. water bath andtotal mixture was incubated for 30 min to facilitate dissolution. WaterTPGS/PSB/Caffeine mixture was mixed on magnetic stiffing plate for 2 hrsat RT. Upon clearing of the mixture, particle size, PSB and caffeinecontent of the resultant emulsion was determined (Table 17b). Theemulsion is transparent but hazy.

TABLE 17b Particle size, PSB content, and Caffeine content of resultantemulsion. PSB Caffeine Manufacturing Particle size by Content, Content,Date Lot# intensity, nm mg/ml Recovery, % mg/ml Recovery, % 2 Aug. 2013005.36.4 42.8 10.79 95 8.56 96

HPLC Analysis. “Pterostilbene 280.M.” method was used for determinationof PSB and caffeine content. PSB content and Caffeine content inresultant emulsion determined by HPLC (“Pterostilbene 280 NM” method)was PSB was 10.79 mg/mL and caffeine 8.56 mg/mL. Recovery is based onthe weighed amount of the pterostilbene/caffeine co-crystal and theweight % provided by the Chromadex C of A. Data indicate that 95% of thePSB used for preparation of this formulation was incorporated into TPGSparticles (Table 17b).

Particle size analysis. The ratio of TPGS to PSB in this emulsion is4.4:1 and this ratio results in a particle size of 42.8 nm which islarger than the particle size in Experiment 16 and again shows that alower ratio of PSB and TPGS result in larger particle sizes. Thepresence of caffeine does not appear to influence the emulsification ofPSB. Three days after manufacture, the filtered emulsion was found tohave crystallized. Data of the supernatant or mother liquors is shown inTable 17 c below. Both PSB and caffeine have come out of solution. Theratio of TPGS to PSB in the supernatant or mother liquors is 9.1:1. Theresults for particle size and PSB and caffeine content of thesupernatant or mother liquors suggests the maximum stable concentrationsof PSB and caffeine and minimum ratio of PSB to TPGS in an emulsion fora given concentration of TPGS (50 mg/mL). The ratio of TPGS to PSB inthe supernatant is again 9.1:1. The emulsion of experiment 17 givessimilar results to that of experiment 16; a critical ratio of 9 to 1 ofTPGS to PSB in the presence of caffeine is necessary to form a stableemulsion of PSB at greater than 0.6 mg/mL PSB.

TABLE 17 c Particle size, PSB content, and particle stability ofresultant emulsion. Particle Days size by PSB Caffeine Manufacturingpast intensity, Content, Recovery, Content, Recovery, Date Lot# mfg Nmmg/ml % mg/ml % 2 Aug. 2013 005.36.4 3* 13.5 5.51 49 4.63 51 *theemulsion crystallized, analysis is of the supernatant

Experiment 18. Preparation of a Cremophor® ELP emulsion. To prepareemulsion containing 5% v/v of Cremophor in distilled water and measureparticle size.

TABLE 18 a Formulation composition. Volume, mL Date Lot# MaterialCremophor Water 27 Aug. 2013 005.39.1 Mixture 1 mL 19 mL

Distilled water, 19 mL, was added to 1 mL of Cremophor in a 20 mL vial,capped and heated to 60° C. then mixed to form a nearly clear solution.

TABLE 18 b Particle size of resultant emulsion. Manufacturing Date Lot#Particle size by intensity, nm 27 Aug. 2013 005.39.1 12.6 28 Aug. 2013005.39.1 12.8

Experiment 19. Preparation of PSB/Cremophor ELP emulsion. To prepareemulsion containing 1.0 mg/ml of PSB and 5 mg/mL Cremophor.

TABLE 19 a Formulation composition. Weighed, mg (per 100 ml) Date Lot#Material Cremophor PSB 25 Aug. 2013 005.38.1 Mixture 5038 1007

5.038 g of Cremophor and 1.007 g of PSB were weighed and placed in 200ml beaker. Cremophor is a viscous oil at RT. The beaker was placed in a65° C. water bath and over 30 minutes the Cremophor dissolved the PSBbased on visual observation. 10 ml of 95% ethanol were added to the oilymixture, and the beaker containing the ethanol mixture of Cremophor/PSBwas placed in the water bath, and incubated at 60° C. for 20 min untilthe oil dissolved, and ethanol was evaporated with a Nitrogen stream.The residual ethanol Cremophor/PSB mixture was placed into vacuum ovenovernight.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mlof distilled water was added to the highly viscous clear film ofCremophor/PSB mixture, the beaker was placed to 60° C. water bath andthe total mixture was incubated for 30 min to facilitate dissolution.The water Cremophor/PSB mixture was mixed on magnetic stiffing plate for1 hr at RT. The mixture never cleared and retained a milky appearance.Particle size of the resultant suspension was determined immediately andthe following day (Table 19b). No settling or crystallization at daysone and three.

TABLE 19 b Particle size of resultant suspension. Manufacturing DateLot# Particle size by intensity, nm 27 Aug. 2013 005.38.1 286.9 28 Aug.2013 005.38.1 339.3

Particle size analysis. The ratio of Cremophor to PSB in this emulsionis 5:1. This ratio resulted in a particle size of approximately 300 nm.

Experiment 20. Preparation of PSB/Caffeine/Cremophor emulsion. Toprepare emulsion containing 10 mg/ml of PSB from the PSB/Caffeineco-crystal (56.2% w/w PSB and 45.0% w/w Caffeine) with Cremophor.

TABLE 20 a Formulation composition. Weighed, mg Calculated (per 100 ml)Weight, mg PSB/ based on C of A Date Lot# Material Cremophor caffeinePSB Caffeine 25 Aug. 2013 005.38.2 Mixture 5000* 1773 996 798 *Actualweight not recorded (5 mL)

Cremophor (5 mL) and 1.773 g of PSB/caffeine were weighed and placed in200 ml beaker. 10 ml of 95% ethanol were added to the solids, and thebeaker containing ethanol mixture of Cremophor/PSB/caffeine was placedin the water bath, and incubated at 60° C. for 30 min The solution didnot clear and upon cooling appeared to crystallize. 10 mL of 95% ethanolwas added and incubated at 60° C. for an additional 30 min until thesolution cleared. During evaporation with a nitrogen stream the solutioncrystallized. The beaker with ethanol/Cremophor/PSB/caffeine solids wasplaced into vacuum oven 0/N.

Emulsion preparation: The beaker was removed from vacuum oven and 100 mlof distilled water was added to solids of Cremophor/PSB/Caffeine, thebeaker was placed to 60° C. water bath and total mixture was incubatedfor 30 min to facilitate dissolution. The water/Cremophor/PSB/Caffeinemixture was mixed on magnetic stiffing plate for 1 hr at RT. The mixtureretained a milky appearance. Particle size of the resultant suspensionwas determined in Table 20b. No settling or crystallization noted ondays one and three.

TABLE 20 b Particle size of resultant suspension. Particle size byintensity, Manufacturing Date Lot# nm 27 Aug. 2013 005.38.2 375.6 28Aug. 2013 005.38.2 371.2

Particle size analysis. The ratio of Cremophor to PSB in this emulsionis approximately 5:1. This ratio resulted in a particle size ofapproximately 375 nm and has large particle size.

The present application discloses unique compositions with specificratios of formulation components that produce stable RVT- orPSB-containing nanoparticles in the range from 20 to 40 nm Thenanoparticulate emulsion is stable in a variety of beverages providingthe calculated beneficial dose of resveratrol or pterostilbene, or amixture thereof, per serving. The above compositions are prepared with aRVT- and PSB-containing nanoparticles where the stable, solublecompositions of RVT- and PSB-containing nanoparticles resulted insimilar stable and clear solutions where a mixture of RVT- andPST-containing nanoparticles were prepared.

The foregoing examples of the related art and limitations are intendedto be illustrative and not exclusive. While a number of exemplaryembodiments, aspects and variations have been provided herein, those ofskill in the art will recognize certain modifications, permutations,additions and combinations and certain sub-combinations of theembodiments, aspects and variations. It is intended that the followingclaims are interpreted to include all such modifications, permutations,additions and combinations and certain sub-combinations of theembodiments, aspects and variations are within their scope. The entiredisclosures of all documents cited throughout this application areincorporated herein by reference.

What is claimed: 1-20. (canceled)
 21. A stable and homogeneous aqueousformulation comprising: a) a composition comprising pterostilbene,resveratrol and a rebaudioside; and b) an emulsifying agent in an amountsufficient to solubilize the composition comprising pterostilbene,resveratrol and a rebaudioside to form the stable and homogeneousformulation; wherein the stable and homogeneous formulation isformulated in an aqueous based beverage.
 22. The stable and homogenousaquesous formulation of claim 21, wherein the rebaudioside isrebaudioside A (Reb A).
 23. The stable and homogeneous aqueousformulation of claim 21, wherein the emulsifying agent is selected fromthe group consisting of TPGS, TPGS-300, TPGS-500, TPGS- 600, TPGS-750,TPGS-1000, TPGS-M, TPGS-300-M, TPGS-500-M, TPGS-600-M, TPGS-750- M andTPGS-1000-M, or a mixture thereof.
 24. The stable and homogeneousaqueous formulation of claim 21, wherein the emulsifying agent is TPGS.25. The stable and homogeneous aqueous formulation of claim 21, whereinthe formulation is an emulsion or a clear solution.
 26. The stable andhomogeneous aqueous formulation of claim 21, wherein the solution is inwater.
 27. The stable and homogeneous aqueous formulation of claim 21,wherein the aqueous based beverage is selected from the group consistingof a clear beverage, soda and fruit juice.
 28. The stable andhomogeneous aqueous formulation of claim 27 wherein the aqueous basedbeverage is selected from the group consisting of vitamin water,carbonated water or beverages and non-carbonated water or beverages. 29.The stable and homogeneous aqueous formulation of claim 27 wherein thesoda is selected from Coke® or Pepsi®.
 30. The stable and homogeneousaqueous formulation of claim 27 wherein the fruit juice is selected fromthe group consisting of orange juice, apple juice, carrot juice andMinute Maid®.
 31. The stable and homogeneous aqueous formulation ofclaim 27 wherein the aqueous based beverage is the vitamin water LifeWater, is Gatorade®, or is PowerAde®.